3. Disc and bone Cells matrix

4. Bone marrow stromal stem cells Skeletal stem cells (SSCs), also known as bone-marrow stromal stem cell, or mesenchymal stem cell

8. Tissue Engineering Triad

9. Core concept

16. Induction of MSCs to Osteoblast

18. Characterization of Osteoblasts

21. TISSUE ENGINEERING APPROACH TO BONE REPAIR BY AUTOLOGOUS BMSCs LOADED ON SUPPORT SCAFFOLDS

22. Topic study cells scaffold The effects of transplantation of osteoblastic cells with bone morphogenetic protein (BMP)/carrier complex on bone repair In vitro and in vivo (Sprague-Dawley Rats) {Bone vol 29, no.2, august 2001:169-75} Osteoblastic/BMP Poly-D,L-lactic-co-glycolic acid/gelatin sponge (PGS) Engineered allogeinic mesenchymal stem cells repair femoral segmental defect in rats Animal study (rat) {J Orthop Res 21 (2003) 44-53} MSC engineered with the gene for BMP-2 Collagen gel (vitrogen 100, 95-98% type 1) Calcium phosphate scaffold and bone marrow for bone reconstruction in irradiated area: a dog study animal model (dog) Bone 36 (2005) 323– 330 Bone marrow macroporous biphasic calcium phosphate bone substitute (MBCPk, Biomatlante, Vigneux de Bretagne, France) Treatment of Long Tubular Bone Defect of Rabbit Using Autologous Cultured Osteoblasts Mixed with Fibrin Animal model (New Zealand white rabbits ) J. of Korean Orthopaedic Society Volume 9, Number 1, May, 2006 autologous cultured osteoblasts fibrin

23. Topic study cells scaffolds Bone Formation of Cultured Osteoblasts in Bone Defect of Radial Shaft of Rabbit Animal model (New Zealand white rabbits ) J. of Korean Orthop. Assoc. 2005; 40: 453-459 autologous cultured osteoblasts The effect of implants loaded with autologous mesenchymal stem cells on the healing of canine segmental bone defects. non-union defect in adult dog f e mor a J. Bone Joint Surg. Am.1998; 80, 985–996. Autologous MSC hydroxyapatite: beta-tricalcium phosphate (65:35) Autologous bone marrow stromal cells loaded onto porous hydroxyapatite ceramic accelerate bone repair in critical-size defects of sheep long bones. full-thickness gaps of tibia diaphysis in adult sheep J. Biomed. Mater. Res. 2000;49, 328–337. Autologous bone marrow stromal cells bioceramic composites Transplantation of marrow-derived mesenchymal stem cells and platelet-rich plasma during distraction osteogenesis—a preliminary result of three cases Clinical study achondroplasia patients bilaterally, and three femoral and one tibial lengthenings were done in four patients because of a limb length discrepancy, which was secondary to trauma (two limbs), congenital pseudarthrosis of the tibia (one), and developmental coxa vara (one) Bone 35 (2004) 892– 898 osteoblast-like cells platelet-rich plasma (PRP)

25. Topic study cells scaffolds Enhanced Tibial Osteotomy Healing with Use of Bone Grafts Supplemented with Platelet Gel or Platelet Gel and Bone Marrow Stromal Cells A prospective, randomized, controlled study . Thirty-three patients undergoing high tibial osteotomy to treat genu varum . J Bone J Surg.Br.2007 ; 11 Osteoblast Platelet gel

26. Topic study cells scaffolds Treatment of Osteonecrosis of the Femoral Head with Implantation of Autologous Bone-Marrow Cells prospective cohort study. T hirteen patients (eighteen hips) with stage-I or II osteonecrosis of the femoral head . J Bone J Surg.Am. 2004 autologous bone-marrow mononuclear cells

27. Study by Aziz Nather Department of orthopaedic surgery National University of Singapore

32. Conclusion 􀂄 Addition of MSCs improved the biological healing of the inert allografts. 􀂄 Addition of PRP did not have the same effect. 􀂄

36. Disc Cells matrix Water and proteoglycans increase from outer the annulus to the inner nucleus, and, in contrast, collagens are inversely distributed Collagen type I proteoglycans Nucleus pulposus chondrocyte-like cell Inner annulus Chondrocyte-like cells Outer annulus fibroblast or fibrocyte-like-cells End-plate chondrocyte Proteoglycans : aggrecans, versican, decorin, biglycan, fibromodulin, lumican, and perlecan. Collagen type II

41. M E T H O D S three sequential stages of experiments were designed :

43. Study on : 1. Developing of external compression and distraction device External compression device External distraction device Stage 2 : Interphase stage

45. 3. Establishing of method of isolation, culture, and preparation of transplantation of bone marrow stromal stem cells . Illiac crest bone marrow aspiration from rabbit donor Day 30 x 40

47. In vitro Study : Day 30 : bMSCs embedded in atelocollagen

49. kept alive for 8 weeks 24 new zealand white rabbits , average weight 3.34 kg Group 1 Group 2 Group 4 Group 3 External compression device, 2.3 MPa, loaded into L4-L5 for 14 days External compression device was removed 8 weeks unload sacrificed bMSCs transplantation Distract ion of the intervertebral disc bMSCs and IVD distraction

50. Tissue preparation : 22 days Lateral digital X-ray, Micro-CT scan -> disc height Macroscopic : morphological grade (Thomson, et al) Microscopic : histological score (Masuda and Boos,et al) and proteoglycans grade (Norcross, et al) Cell viability (Tunel reaction) and cell labeling (CFDA)

51. Surgical procedure

53. transplantation of 0.08 ml bMSCs solution are injected into disc by posterolateral approach and image intensifier guided through a 25-gauge syringe.

54. Image intensifier-guided

56. In vivo Cell labeling : 8 weeks after transplantation group 1 (bMSCs) group 4 (distraction and bMSCs)

57. RESULTS DISCUSSIONS

60. Group 3 Group 1 Group 2 Group 4

67. CONCLUSIONS

69. Cell Therapy and Stem cells for Cartilage Defect Courtesy of Dr. Andri Lubis, Sp OT Orthopaedic and Traumatology Division Faculty of Medicine University of Indonesia

71. cartilage MFC denuded 3x5 cm Osteochondral lesion

73. JBJS Am 2006;88:2502-20 Autologous Chondrocyte Implantation (ACI)

74. JBJS Am 2006;88:2502-20

75. ACI Technology Brittberg et al., NEJM 1994; 331:889-95

79. J Kor Arthros Soc 2007;11:1-5

80. Bone Marrow-Derived Mesenchymal Stem Cells

82. Courtesy of Prof. James Hui, National University Hospital, Singapore

85. Courtesy of Prof. James Hui, NUH

86. Radiographics 2007; 27:207-22

87. Eur Radiol 2007; 17:103–18

89. Tissue Engineering and Cell Based Therapies: a 21 st Century Treatment Dr. Caplan

91. The End Thank you for the attention